Objective: Selective progesterone receptor modulators (SPRMs), such as asoprisnil (J867) and ulipristal (CDB-2914), have been shown to reduce fibroid volume in vivo and to induce apoptosis in vitro. CDB-4124 (telapristone), a SPRM with different side groups, also reduced fibroid volume in vivo, and we hypothesized that this SPRM would also cause apoptosis in cultured fibroid cells. Methods: Immortalized, progesterone receptor-positive fibroid cells, known to be capable of apoptosis, were grown to 80% confluence in serum-containing media. Cells were then treated for 48 hours in serum-free media with 0, 10, 100, or 1000 nmol/L CDB-4124. Actinomycin-D and staurosporine were used as positive controls to induce apoptosis. Apoptosis was quantified using a TUNEL-fluorescein kit. Images were captured with a widefield-fluorescence microscope and analyzed using MetaMorph image analysis software. To validate results, Western blots of total cell lysates were probed for cleaved caspase-3 (c-CASP3). Experiments were repeated 3 times using independent cell batches. Results: Analysis of 19 712 nuclei indicated 14.8% ± 10.9% (mean ± SEM), 8.4% ± 4.6%, 8.2% ± 4.7%, and 9.3% ± 6.3% apoptosis in 0, 10, 100, and 1000 nmol/L CDB-4124-treated cells, respectively. There was no evidence of elevated c-CASP3 over vehicle control after treatment with CDB-4124. Conclusion: CDB-4124 did not significantly induce apoptosis in cultured fibroid cells under the conditions described suggesting apoptosis may not be the main pathway responsible for CDB-4124-induced fibroid shrinkage. Variations in SPRM biological effects may be due to differences in fibroid source cells, binding kinetics, or extracellular matrix characteristics, and can be exploited in further investigations of the mechanisms of action of SPRMs in fibroid biology.
LeppertPCBaginskiTPrupasCCatherinoWHPletcherSSegarsJH. Comparative ultrastructure of collagen fibrils in uterine leiomyomas and normal myometrium. Fertil Steril. 2004;82(3):1182–1187.
2.
Day BairdDDunsonDBHillMCCousinsDSchectmanJM. High cumulative incidence of uterine leiomyoma in black and white women: ultrasound evidence. Am J Obstet Gynecol. 2003;188(1):100–107.
3.
FriedmanAJHarrison-AtlasDBarbieriRLBenacerrafBGleasonRSchiffI. A randomized, placebo-controlled, double-blind study evaluating the efficacy of leuprolide acetate depot in the treatment of uterine leiomyomata. Fertil Steril. 1989;51(2):251–256.
4.
ReinMS. Advances in uterine leiomyoma research: the progesterone hypothesis. Environ Health Perspect. 2000;108(suppl 5):791–793.
5.
CarrBRMarshburnPBWeatherallPT. An evaluation of the effect of gonadotropin-releasing hormone analogs and medroxyprogesterone acetate on uterine leiomyomata volume by magnetic resonance imaging: a prospective, randomized, double blind, placebo-controlled, crossover trial. J Clin Endocrinol Metab. 1993;76(5):1217–1223.
6.
PintiauxAChabbert-BuffetNFoidartJM. Gynaecological uses of a new class of steroids: the selective progesterone receptor modulators. Gynecol Endocrinol. 2009;25(2):67–73.
7.
EngmanMGranbergSWilliamsARMengCXLalitkumarPGGemzell-DanielssonK. Mifepristone for treatment of uterine leiomyoma. A prospective randomized placebo controlled trial. Hum Reprod. 2009;24(8):1870–1879.
8.
ChwaliszKLarsenLMattia-GoldbergCEdmondsAElgerWWinkelCA. A randomized, controlled trial of asoprisnil, a novel selective progesterone receptor modulator, in women with uterine leiomyomata. Fertil Steril. 2007;87(6):1399–1412.
9.
ChenWOharaNWangJ. A novel selective progesterone receptor modulator asoprisnil (J867) inhibits proliferation and induces apoptosis in cultured human uterine leiomyoma cells in the absence of comparable effects on myometrial cells. J Clin Endocrinol Metab. 2006;91(4):1296–304.
10.
OharaNMorikawaAChenW. Comparative effects of SPRM asoprisnil (J867) on proliferation, apoptosis, and the expression of growth factors in cultured uterine leiomyoma cells and normal myometrial cells. Reprod Sci. 2007;14(8 suppl):20–27.
11.
LevensEDPotlog-NahariCArmstrongAY. CDB-2914 for uterine leiomyomata treatment: a randomized controlled trial. Obstet Gynecol. 2008;111(5):1129–1136.
12.
XuQTakekidaSOharaN. Progesterone receptor modulator CDB-2914 down-regulates proliferative cell nuclear antigen and Bcl-2 protein expression and up-regulates caspase-3 and poly(adenosine 5'-diphosphate-ribose) polymerase expression in cultured human uterine leiomyoma cells. J Clin Endocrinol Metab. 2005;90(2):953–961.
13.
AttardiBJBurgensonJHildSAReelJRBlyeRP. CDB-4124 and its putative monodemethylated metabolite, CDB-4453, are potent antiprogestins with reduced antiglucocorticoid activity: in vitro comparison to mifepristone and CDB-2914. Mol Cell Endocrinol. 2002;188(1-2):111–123.
14.
WiehleRBGoldbergJBrodniewiczTDziedzicKJJabiry-ZieniewiczZ. Effects of a new progesterone receptor modulator, CDB-4124, on fibroid size and uterine bleeding. (US obstetrics and Gynecology)Touch Briefings. 2008;3(1):17–20.
15.
IoffeOBZainoRJMutterGL. Endometrial changes from short-term therapy with CDB-4124, a selective progesterone receptor modulator. Mod Pathol. 2009;22(3):450–459.
16.
MalikMWebbJCatherinoWH. Retinoic acid treatment of human leiomyoma cells transformed the cell phenotype to one strongly resembling myometrial cells. Clin Endocrinol (Oxf). 2008;69(3):462–470.
17.
MalikMMendozaMPaysonMCatherinoW. Curcumin, a nutritional supplement with antineoplastic activity, enhances leiomyoma cell apoptosis and decreases fibronectin expression. Fertil Steril. 2009;91(5):2177–2184.
18.
CouldwellWTHintonDRHeS. Protein kinase C inhibitors induce apoptosis in human malignant glioma cell lines. FEBS Lett. 1994;345(1):43–46.
19.
JeyasuriaPWetzelJBradleyMSubediKCondonJ. Progesterone-regulated caspase 3 action in the mouse may play a role in uterine quiescence during pregnancy through fragmentation of uterine myocyte contractile proteins. Biol Reprod. 2009;80(5):928–934.
20.
WadeHKobayashiSEatonM. Multimodal regulation of E2F1 gene expression by progestins. Mol Cell Biol. 2010;30(8):1866–1877.
21.
MalikMCatherinoWH. Novel method to characterize primary cultures of leiomyoma and myometrium with the use of confirmatory biomarker gene arrays. Fertil Steril. 2007;87(5):116–172.
22.
CatherinoWHMalikMDriggersPChappelSSegarsJDavisJ. Novel, orally active selective progesterone receptor modulator CP8947 inhibits leiomyoma cell proliferation without adversely affecting endometrium or myometrium. J Steroid Biochem Mol Biol. 2010;122(4):279–286.
23.
LuoXYinPCoonVJChengYHWiehleRDBulunSE. The selective progesterone receptor modulator CDB4124 inhibits proliferation and induces apoptosis in uterine leiomyoma cells. Fertil Steril. 2010;93(8):2668–2673.
24.
SinghPKrishnaA. Effects of GnRH agonist treatment on steroidogenesis and folliculogenesis in the ovary of cyclic mice. J Ovarian Res. 2010;3:26.
25.
MiuraSKhanKNKitajimaM. Differential infiltration of macrophages and prostaglandin production by different uterine leiomyomas. Hum Reprod. 2006;21(10):2545–2554.
26.
ReinMSPowellWLWaltersFC. Cytogenetic abnormalities in uterine myomas are associated with myoma size. Mol Hum Reprod. 1998;4(1):83–86.
27.
LobelMSomasundaramPMortonC. The genetic heterogeneity of uterine leiomyomata. Obstet Gyneco Clin North Am. 2006;33(1):13–39.
28.
HodgeJQuadeBRubinM. Molecular and cytogenetic characterization of plexiform leiomyomata provide further evidence for genetic heterogeneity underlying uterine fibroids. Am J Pathol. 2008;172(5):1403–1410.
29.
YinPLinZChengY. Progesterone receptor regulates Bcl-2 gene expression through direct binding to its promoter region in uterine leiomyoma cells. J Clin Endocrinol Metab. 2007;92(111):4459–466.
30.
MorikawaAOharaNXuQ. Selective progesterone receptor modulator asoprisnil down-regulates collagen synthesis in cultured human uterine leiomyoma cells through up-regulating extracellular matrix metalloproteinase inducer. Hum Reprod. 2008;23(4):944–951.
31.
XuQOharaNLiuJ. Progesterone receptor modulator CDB-2914 induces extracellular matrix metalloproteinase inducer in cultured human uterine leiomyoma cells. Mol Hum Reprod. 2008;14(3):181–191.
32.
XuQOharaNChenW. Progesterone receptor modulator CDB-2914 down-regulates vascular endothelial growth factor, adrenomedullin and their receptors and modulates progesterone receptor content in cultured human uterine leiomyoma cells. Hum Reprod. 2006;21(9):2408–2416.
